OBJECTIVE:Evidence of increased apoptosis is observed in periodontitis, and may be associated with destruction of the periodontal tissue caused by the increased cell death, with the release of danger signals and subsequent stimulation of the proinflammatory processes. However, the exact mechanisms associated with these processes remain unclear. This study aimed to investigate the presence of the periodontal pathogen Treponema denticola, apoptosis, High Mobility Group Box1 as a damage-associated molecular pattern and several inflammatory markers in periodontitis and gingivitis subjects. MATERIALS AND METHODS:Soft tissue specimens from gingival tissues of periodontitis and gingivitis patients were used for immunohistochemical and immunofluorescence staining of Treponema denticola chymotrypsin-like proteinase (CTLP), apoptosis markers, High Mobility Group Box1, Toll-like receptor 4, inflammatory cell markers and proinflammatory cytokines. RESULTS:Treponema denticola was detected in all periodontitis-affected tissues. This was associated with a significant increase in the number of apoptotic cells, including macrophages, alterations in the expression of High Mobility Group Box1 and its receptor, and increased levels of proinflammatory cytokines compared with gingivitis. CONCLUSIONS:In summary, the presence of Treponema denticola (especially its CTLP), apoptosis, High Mobility Group Box1, and inflammatory markers suggest their potential involvement in the pathogenesis of periodontitis.

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Background: Periodontitis causes an increased receptor activator level in the nuclear factor-κβ ligand (RANKL), which is one of the inflammatory mediators that plays a role in osteoclastogenesis. The open flap debridement (OFD) technique is the preferred treatment when accompanied by regenerative periodontal treatment using guided tissue regeneration (GTR) and guided bone regeneration (GBR). Carbonated hydroxyapatite is a GBR material that serves as a scaffold and has strong osteoconductive properties for bone regeneration. Propolis is natural product that can decrease osteoclastogenesis in periodontitis by decreasing the RANKL expression. Purpose: To investigate the RANKL expression after open flap debridement by applying carbonated hydroxyapatite to 10% propolis in the alveolar bone of rabbits. Methods: Nine induced-periodontitis rabbits (Oryctolagus cuniculus) were divided into three treatment groups of Group A OFD, Group B OFD followed by the application of carbonated hydroxyapatite, and Group C OFD followed by application of 10% propolis-carbonated hydroxyapatite. Each group was selected one to euthanised on the seventh, 14th and 28th day, respectively, and prepared using histology slides. The data was analysed using a two-way ANOVA followed by a post-hoc LSD test (p<0.05). Results: The RANKL expression in each group showed significant differences (p=0.00; p<0.05) on the seventh, 14th and 28th day. The post-hoc LSD test showed that the RANKL expression in the treatment group with carbonated hydroxyapatite-10% propolis had significant differences (p<0.05) in the intergroup analysis at different time points. Conclusion: Combining 10% propolis with carbonated hydroxyapatite in OFD treatment can decrease the RANKL expression in a rabbit’s alveolar bone.

Carbonated hydroxyapatite is ideal as a bone graft material because it has similar organic matters to the bone, excellent osteoconductive properties, and good biodegradation in the body. Hydroxyapatite contains the risk of being contaminated by bacteria called Porphyromonas gingivalis (P. gingivalis) in the oral cavity because it has no vascularization, therefore, facilitating adhesion of bacteria, and when applied in the oral cavity, it may cause an infection that then inhibits healing. Thus, it is necessary to use a material that has an antibacterial effect with low potential of causing resistance to treat the postsurgical infection properly. Propolis has antibacterial, antiviral, antifungal, antitumor, and immunomodulatory activities. Propolis contains a large number of flavonoids and phenols. The phenol compound in propolis is usually called caffeic acid phenethyl ester (CAPE), and it has a good antibacterial property. The study aims to evaluate the antibacterial effect of carbonated hydroxyapatite when immersed with different propolis concentrations of 2.5%, 5%, 7.5%, and 10% for 24 h and to measure the zone of inhibition against P. gingivalis. The Kruskal–Wallis test resulted in p = 0.00 (p < 0.05), indicating that there were significant differences among the test groups. The data processing was followed by Mann–Whitney U-test, and the results showed a significant difference in the group of carbonated hydroxyapatite-10 % propolis compared with the other groups. Inhibition zone of carbonated hydroxyapatite that immersed with propolis 10% showed the largest mean of diameters zone of inhibition.